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Fig. 1 | Biomarker Research

Fig. 1

From: Analytical validation and initial clinical testing of quantitative microscopic evaluation for PD-L1 and HLA I expression on circulating tumor cells from patients with non-small cell lung cancer

Fig. 1

Overview of ESP-Quantitative Microscopy. A The ExtractMax automated platform employs ESP technology to capture and stain CTCs. B Glass coverslips under adhesive silicone isolators facilitate high-quality imaging of suspended cells in a total volume of 100 uL across a total area of ~ 7 × 7 mm. C Focus-controlled imaging accounts for variability in the focal plane due to tilt in the microscope stage across the entire imaging area. D The focus surface is generated by registering a Z-plane offset at each corner of the imaging area (as indicated by the heat map), enabling automated Z-plane adjustment during the acquisition of individual small image tiles (~1um each) of high magnification (10x) acquired in a grid across the entire imaging area. E Biomarker intensity is quantified via the generation of binary layers on the intensity of Hoechst (the 350 channel) after intensity flattening with rolling ball background subtraction using a fully automated image analysis coded “macro”

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Biomarker Research

ISSN: 2050-7771

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